RESUMEN
Myxobacteria use soluble and cell-contact signals during their starvation-induced formation of fruiting bodies. These signals coordinate developmental gene expression with the cell movements that build fruiting bodies. Early in development, the quorum-sensing A-signal in Myxococcus xanthus helps to assess starvation and induce the first stage of aggregation. Later, the morphogenetic C-signal helps to pattern cell movement and shape the fruiting body. C-signal is a 17-kDa cell surface protein that signals by contact between the ends of two cells. The number of C-signal molecules per cell rises 100-fold from the beginning of fruiting body development to the end, when spores are formed. Traveling waves, streams, and sporulation have increasing thresholds for C-signal activity, and this progression ensures that spores form inside fruiting bodies.
Asunto(s)
Myxococcus xanthus/fisiología , Transducción de Señal/fisiología , Stigmatella aurantiaca/fisiología , Myxococcus xanthus/genética , Myxococcus xanthus/crecimiento & desarrollo , Transducción de Señal/genética , Esporas Bacterianas/fisiología , Stigmatella aurantiaca/genética , Stigmatella aurantiaca/crecimiento & desarrolloRESUMEN
Transcriptional factor CarD is the only reported prokaryotic analog of eukaryotic high-mobility-group A (HMGA) proteins, in that it has contiguous acidic and AT hook DNA-binding segments and multifunctional roles in Myxococcus xanthus carotenogenesis and fruiting body formation. HMGA proteins are small, randomly structured, nonhistone, nuclear architectural factors that remodel DNA and chromatin structure. Here we report on a second AT hook protein, CarD(Sa), that is very similar to CarD and that occurs in the bacterium Stigmatella aurantiaca. CarD(Sa) has a C-terminal HMGA-like domain with three AT hooks and a highly acidic adjacent region with one predicted casein kinase II (CKII) phosphorylation site, compared to the four AT hooks and five CKII sites in CarD. Both proteins have a nearly identical 180-residue N-terminal segment that is absent in HMGA proteins. In vitro, CarD(Sa) exhibits the specific minor-groove binding to appropriately spaced AT-rich DNA that is characteristic of CarD or HMGA proteins, and it is also phosphorylated by CKII. In vivo, CarD(Sa) or a variant without the single CKII phosphorylation site can replace CarD in M. xanthus carotenogenesis and fruiting body formation. These two cellular processes absolutely require that the highly conserved N-terminal domain be present. Thus, three AT hooks are sufficient, the N-terminal domain is essential, and phosphorylation in the acidic region by a CKII-type kinase can be dispensed with for CarD function in M. xanthus carotenogenesis and fruiting body development. Whereas a number of hypothetical proteins homologous to the N-terminal region occur in a diverse array of bacterial species, eukaryotic HMGA-type domains appear to be confined primarily to myxobacteria.
Asunto(s)
Proteínas Bacterianas , Stigmatella aurantiaca/metabolismo , Transactivadores/metabolismo , Secuencias AT-Hook/genética , Secuencia de Aminoácidos , Southern Blotting , Quinasa de la Caseína II , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Eliminación de Gen , Prueba de Complementación Genética , Datos de Secuencia Molecular , Myxococcus xanthus/genética , Fosforilación , Proteínas Serina-Treonina Quinasas/metabolismo , Alineación de Secuencia , Especificidad de la Especie , Stigmatella aurantiaca/genética , Stigmatella aurantiaca/crecimiento & desarrollo , Transactivadores/biosíntesis , Transactivadores/genéticaRESUMEN
Alternative sigma factors have been detected in the myxobacterium Stigmatella aurantiaca during indole-induced sporulation, fruiting body formation and heat shock using an antiserum raised against sigma factor SigB. The time course of sigB gene expression was analysed by RT-PCR and by determining beta-galactosidase activity during development in a merodiploid strain that harboured a sigB-lacZ fusion gene. Inactivation of the sigB gene by insertion of the neo gene resulted in the loss of one sigma factor as shown by Western analysis. Neither fruiting body formation nor sporulation, nor the production of possible SigB targets, such as DnaK, GroEL or HspA, were affected.
Asunto(s)
Proteínas Bacterianas/biosíntesis , Regulación Bacteriana de la Expresión Génica , Calor , Factor sigma/biosíntesis , Esporas Bacterianas/metabolismo , Stigmatella aurantiaca/metabolismo , Proteínas Bacterianas/genética , Secuencia de Bases , Proteínas de Choque Térmico/metabolismo , Datos de Secuencia Molecular , Mutación , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factor sigma/genética , Esporas Bacterianas/genética , Stigmatella aurantiaca/genética , Stigmatella aurantiaca/crecimiento & desarrollo , Transcripción GenéticaRESUMEN
3-Deoxy-D-arabino-heptulosonate-7-phosphate (DAHP) synthases catalyse the first step of the shikimate pathway. Two unrelated DAHP synthase types have been described in plants and bacteria. Two type II (aroA(A2) and aroA(A5)) and one type I DAHP synthase gene (aroA001) were identified from the myxobacterium Stigmatella aurantiaca Sg a15. Inactivation of aroA(A5) leads to a mutant that is impaired in the biosynthesis of aurachins, which are electron transport inhibitors and contain an anthranilate moiety. Feeding of anthranilic acid to the mutant culture restores production of aurachins. Inactivation of aroA(A2) and aroA001 does not impair production of aurachins or other known secondary metabolites of S. aurantiaca Sg a15.
Asunto(s)
3-Desoxi-7-Fosfoheptulonato Sintasa/genética , Genes Bacterianos , Quinolonas/metabolismo , Stigmatella aurantiaca/enzimología , Stigmatella aurantiaca/genética , 3-Desoxi-7-Fosfoheptulonato Sintasa/química , 3-Desoxi-7-Fosfoheptulonato Sintasa/metabolismo , Secuencia de Aminoácidos , Clonación Molecular , Cósmidos , Datos de Secuencia Molecular , Familia de Multigenes , Mutación , Alineación de Secuencia , Análisis de Secuencia de ADN , Stigmatella aurantiaca/crecimiento & desarrolloRESUMEN
The myxobacterium Stigmatella aurantiaca is a prokaryotic model used to study intercellular signalling and the genetic determination of morphogenesis. Signalling factors and genes required for the generation of the elaborate multicellular fruiting body are to be identified. Recently, the structure of stigmolone, which is the pheromone necessary for fruiting body formation, was elucidated, and genes involved in development were characterised. Progress has also been made in the genetic accessibility of S. aurantiaca.